We report our success in simultaneously tracking tissue formation and single-cell migration in a mouse embryo on embryonic day 5.5 for 12 h. The microscope, incubator, and observation protocols were comprehensively specialized and optimized to achieve this. The present system revealed new phenomena that had not previously been observed and promises to elucidate the mechanism of mouse embryonic development in the future.
Go Shioi, Tomonobu M Watanabe, Junichi Kaneshiro, Yusuke Azuma, Shuichi Onami (2025) Trans-scale live-imaging of an E5.5 mouse embryo using incubator-type biaxial light-sheet microscopy., Life science alliance, Volume 8, Number 3
Published in 2025 Mar (Electronic publication in Jan. 15, 2025, midnight )
(Abstract) During mouse embryonic development, the embryonic day (E) 5.5 stage represents a crucial period for the formation of the primitive body axis, where the symmetry breaking of cellular states influences the multicellular system. Elucidating the detailed mechanisms of this process necessitates a trans-layered dynamic observation of the embryo and all internal cells. In this report, we present our success in achieving in-toto single-cell observation in a whole hemisphere of an E5.5 embryo for 12 h, using a newly developed incubator-type biaxial light-sheet microscope. To achieve the success, we optimized our microscope system, including an incubator for culture stability, and refining the observation protocol to reduce phototoxicity. Our key discovery is that the scan speed during light-sheet formation plays a critical role in reducing phototoxicity, rather than the irradiation intensity or the interval time between frames. This innovative system not only enabled in-toto single-cell tracking but also led to the discovery of the abrupt shrinking of embryos whose contractile center was located at the extraembryonic ectoderm during monotonous growth up to the E6.5 stage.(MeSH Terms)